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Flower-bud formation suppressor gene and early flowering plant

a technology of suppressor gene and flowering plant, which is applied in the field of new flowering plant suppressor gene, can solve the problems of long time for woody plant progress, and long period of time for woody plant improvemen

Inactive Publication Date: 2007-08-02
NAT AGRI RES ORG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about isolating a gene that controls flower-bud formation in perennial plants. By using this gene or an antisense DNA of it, researchers have been able to create plants with an early flowering property. The invention also includes an isolated protein and a nucleic acid that can be used to regulate the flowering time of plants. The transformation of plants with the antisense nucleic acid resulted in a perennial tree with an early flowering property. Overall, the invention provides a way to control flowering in plants and create early-blooming varieties.

Problems solved by technology

Unlike for herbaceous plants, it takes a long time for woody plants to progress from flowering to seed-setting, that is, woody plants have long juvenile periods, Hence, improvement through breeding of a perennial fruit tree crop, such as Malus (apple) belonging to the family Rosaceae, requires a long period of time.
Thus, juvenility, the unique feature of woody plants, is a significant factor that impedes the efficiency of cross breeding.
However, no method can be said to have exerted any landmark effect (Pinero, M. et al., Plant Physiol.
So it has not been easy to clarify the genes and mechanisms involved in the flowering (Weigel, D. et al., Nature 1995, 377: 495-500; Mandel, M. A. et al., Nature 1995, 377: 522-524).

Method used

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  • Flower-bud formation suppressor gene and early flowering plant
  • Flower-bud formation suppressor gene and early flowering plant
  • Flower-bud formation suppressor gene and early flowering plant

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation of MdTFL Gene

(1) Apple Plant Employed in the Following Experiments

[0121] 15 to 16-year old Jonathan apples (Malus x domestics cv. Jonathan) grafted to Maruba rootstocks were used.

(2) Preparation of poly (A)+ RNA

[0122] Before and after floral initiation stage (differentiation of flower-bud), that is, from July to September, the shoot apices of the plant of (1) above were collected, and then total RNA was prepared by the CTAB method. Specifically, 3 g of frozen apple shoot apex was crushed into a powdery form, suspended in a 3 mL of 2×CTAB solution (2% CTAB, 0.1 M Tris-HCl, pH 9.5, 20 mM EDTA, 1.4 M NaCl, 1% 2-propanol), and then incubated at 65° C. for 10 minutes. The solution was subjected twice to extraction using chloroform isoamyl, added with 1 / 4 volume of 10 M lithium chloride, and then allowed to stand at −20° C. for 2 hours to obtain precipitated RNA. Subsequently, centrifugation is performed at 12,000×g for 10 minutes 4° C. to collect the precipitated RNA.

[01...

example 2

Southern Blot Analysis of MdTFL Gene

(1) Preparation of Genomic DNA

[0137] From the leaves of an apple plant “Jonathan apple,” 1 genomic DNA was prepared by a CTAB method. Specifically, 3 g of apple leaves were frozen in liquid nitrogen, and then quickly crushed in mortar to obtain a powder form. To remove sugars, an extraction buffer (10% polyethylene glycol 6000, 0.35 M sorbitol, 0.1 M Tris-HCl pH 7.5, 1% 2-mercaptoethanol) was added and mixed well in a 50 mL tube. Then, centrifugation was performed at 12,000×g for 5 minutes at room temperature. The supernatant was discarded, and then to the pellet 9 mL of a lytic buffer (0.35 M sorbitol, 0.1 M Tris-HCl pH 7.5, 1% 2-mercaptoethanol) and 1 mL of 10% sarcosine were added, followed by gentle agitation at room temperature for 10 minutes. Then, 10 mL of 2×CTAB (2% CTAB, 0.1 M Tris-HCl pH 9.5, 20 mM EDTA, 1.4 M NaCl, 1% 2-mercaptoethanol) was added, and then the solution was gently shaken at 56° C. for 20 minutes.

[0138] After the abov...

example 3

Northern Blot Analysis of MdTFL Gene

(1) Preparation of RNA

[0142] Total RNA was prepared by the method described in Example 1 (2) above from each apple plant organs including calices, flower petals, stamens, pistils, shoot apices, leaves, cotyledons, stems and roots. In addition, shoot apices and flower-bud portions of the apple plants were collected from June to April of the following year at preset intervals, and then total RNAs of the shoot apices of the flower buds collected at each time were prepared.

(2) Hybridization

[0143] 10 μg of the total RNA obtained in (1) above was subjected to formaldehyde denaturation 1.2% agarose gel electrophoresis, and then transferred to a nylon membrane. The membrane was immersed in a pre-hybridization solution (5×SSC, 10× Denhardt's solution, 10 mM Na2PO4 (pH 6.5), 0.5% SDS, 50% formamide, 10 mg / mL salmon sperm DNA) at 65° C. for 1 hour to perform pre-hybridization. Subsequently, hybridization was performed using an RNA probe that had been l...

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Abstract

According to the present invention, a plant having an early flowering property is provided. Specifically, the present invention provides a transformed plant having a gene that suppresses flower-bud formation or the antisense DNA of this gene.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims a priority from Japanese Patent Application No. 2002-180289 filed Jun. 20, 2002, the disclosure of which is incorporated herein by reference. BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The present invention relates to a novel flower-bud formation suppressor gene which is useful to confer an early flowering property on a perennial plant. The present invention also relates to a recombinant vector which comprises a DNA encoding the above gene or an antisense DNA of the gene, a transformant which comprises the recombinant vector, and seeds obtained from the transformant. The present invention further relates to a method for conferring an early flowering property on a plant, and a method for producing an early flowering plant. [0004] 2. Description of the Related Art [0005] Unlike for herbaceous plants, it takes a long time for woody plants to progress from flowering to seed-setting, that is, w...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01H1/00C12N15/82A01H5/00C07K14/415C12N5/04C12N5/10C12N15/09C12N15/29
CPCC07K14/415C12N2510/00C12N15/827C12N5/04
Inventor KOTODA, NOBUHIROWADA, MASATOSOEJIMA, JUNICHI
Owner NAT AGRI RES ORG
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